中国禽(番鸭)呼肠孤病毒YB株S4基因序列分析

Cloning and cDNA sequence analysis of the S4 gene of the avian(muscovy duck) reovirus isolate(DRV-YB) in China

应用非免疫番鸭胚增殖中国禽(番鸭)呼肠孤病毒YB分离株,用LS TRIAZOL提取病毒RNA,反转录-聚合酶链反应(RT-PCR)扩增中国禽(番鸭)呼肠孤病毒YB分离株S4基因节段cDNA。将S4cDNA克隆到PMD18-T载体上,并进行了鉴定和核苷酸序列测定。序列分析结果,克隆的S4cDNA共1 124 bp,包括非编码区和完整的阅读框架。分子进化系统分析表明该毒株与禽呼肠孤病毒(鸡呼肠孤病毒)的亲缘关系较远,DRV-YB与DRV-89330同缘率为93.3%。

Muscovy duck reovirus YB（DRV-YB） China strain was grown in 7-day-old embryonated muscovy duck eggs unaffectedly.Virion RNA was isolated by treatment of aliquots of the virus with IS TRIAZOL. The cDNA of the S4 gene frame of DRV-YB was amplified by reverse transcription-polymerase chain reaction, cloned into pMDT-18 vector and sequenced. The results show that the cloned 1 124 bp $4 gene, including the complete open reading frames （ORFs）. Evolution analysis indicated the isolate YB was in far relation to avian reovirus strains （chicken reovirus strains）, and nucleotide homogeneity of S4 between isolate YB in China and muscovy duck reovirus strains in France （DRV-89330） was 93.3 %.